Objective To explore the age-related biological properties of bone-derived mesenchymal stem cells(BMSCs)from mice of different age groups and their osteogenic differentiation induced by bone morphogenetic protein 2(BMP2).Methods Eight C57BL/6J mice were divided into a young group(4 weeks old,weighing 10～15 g,n=4)and an old group(12 months old,weighing 20～25 g,n=4),with half male and half female.MSCs were extracted from the whole bones of the 2 groups of mice.After the obtained cells were identified with flow cytometry for the surface markers,β-galactosidase staining was employed to compare the senescence level of BMSCs,MTT and EdU incorporation assays were conducted to compare the proliferation and self-renewal abilities of between the 2 groups.Western blotting was employed to analyze the expression of CyclinD1 and P21 in BMSCs.Then ALP staining,Alizarin Red staining and RT-qPCR were used to evaluate the osteogenic differentiation ability of the cells.RNA sequencing was performed to compare the differential gene expression in BMP2-induced BMSCs.Lastly,the sequencing results were re-confirmed by using flow cytometry.Results Flow cytometry showed that the sorted and cultured mouse BMSCs met the criteria for MSCs.The results of β-galactosidase staining indicated that the senescence level of BMSCs in the old group was significantly higher than that in the young group(P＜0.05).MTT and EdU doping experiments revealed that the cell viability and proliferation ability of BMSCs were significantly lower in the old group than the young group(P＜0.05).Western blotting displayed that the expression level of cell cycle protein CyclinD1 was lower,whereas that of cell cycle inhibitory factor P21 was significantly higher in the BMSCs from the old group than the cells from the young group(P＜0.05).ALP/Alizarin Red staining and RT-qPCR demonstrated that the BMSCs from the young group had stronger osteogenic differentiation capacity after BMP2 treatment when compared the cells of the old group(P＜0.05).RNA sequencing results displayed that the changing profile of CD51 expression was in opposite trends in the young and old BMSCs after BMP2 treatment.Finally,flow cytometry revealed that the percentage of CD51+cells within the CD45-cells was significantly higher in the young group than the old group.Conclusion The decrease in the percentage of CD51+cells among CD45-cells in aged BMSCs is closely associated with their decreased responsiveness to BMP2-induced osteogenic differentiation.

Animals ; SARS-CoV-2 ; Antibodies, Neutralizing ; Macaca mulatta ; COVID-19/prevention & control* ; Antibodies, Viral ; Vaccines

Objective:To investigate effects of the autophagy inducer rapamycin and autophagy inhibitor 3-methyladenine on viral structures and biosynthesis of functional proteins in dorsal root ganglia in a guinea pig model of varicella-zoster virus (VZV) infection, and to explore their possible mechanisms.Methods:VZV was cultured and proliferated in human embryonic lung fibroblasts (HELFs) , and peripheral blood mononuclear cells (PBMCs) were isolated from guinea pigs. VZV-HELFs and PBMCs were co-cultured for 18-20 hours, and VZV-PBMCs were collected by centrifugation. Thirty-two guinea pigs were randomly and equally divided into 4 groups (8 mice in each group) : blank control group was injected with autologous PBMCs via the medial canthal venous plexus; autophagy inhibition group, autophagy induction group, and VZV infection group were intraperitoneally injected with 3 mg/kg 3-methyladenine solution, 0.5 mg/kg rapamycin solution, and the same volume of 0.9% NaCl solution respectively, followed 2 hours later by injections with 50 μl of VZV-PBMCs via the medial canthal venous plexus. Fourteen days later, the guinea pigs in each group were sacrificed, and dorsal root ganglion tissues were collected. The transmission electron microscope was used to observe the morphology of virus particles, as well as the morphology and number of autophagic vesicles, Western blot analysis was performed to determine the expression of VZV nucleocapsid protein (NCP) , immediate-early protein 62 (IE62) , and autophagy-related proteins Beclin-1 and p62, and immunohistochemical study to determine the expression of anti-VZV antibodies in VZV-infected dorsal root ganglia. Statistical analysis was carried out by using two-independent-sample t test, one-way analysis of variance, least significant difference- t test or Kruskal-Wallis H test. Results:Nucleocapsid-containing virions and scattered autophagosomes were seen in the dorsal root ganglia in the VZV infection group under the transmission electron microscope. The number of autophagic vesicles significantly differed among the blank control group, VZV infection group, autophagy induction group and autophagy inhibition group ( M[ Q1, Q3]: 0, 5[4, 6], 7[5, 9], 0, respectively; H = 135.60, P < 0.01) , and was significantly higher in the VZV infection group than in the blank control group and autophagy inhibition group (both P < 0.05) , as well as in the autophagy induction group than in the autophagy inhibition group ( P<0.05) , but there was no significant difference between the VZV infection group and autophagy induction group ( P>0.05) . Western blot analysis showed that the expression level of IE62 protein was significantly higher in the VZV infection group (1.49 ± 0.06) than in the blank control group (0.50 ± 0.09, t = 9.17, P < 0.05) ; the expression of anti-VZV antibodies was significantly lower in the autophagy inhibition group than in the autophagy induction group and VZV infection group ( t = 9.24, 7.78, respectively, both P < 0.01) , while there was no significant difference between the autophagy induction group and VZV infection group ( P > 0.05) . Conclusion:Autophagy occurred in the dorsal root ganglia of guinea pigs after VZV infection; the inhibition of autophagy could affect the structure of VZV and decrease the expression of VZV functional proteins in the dorsal root ganglia of guinea pigs.

Objective: To explore the efficacy of an individualized cardiac rehabilitation education model on the knowledge level of cardiac rehabilitation and cardiac rehabilitation exercise for patients after percutaneous coronary intervention (PCI). Methods: This study adopted the method of quasi-experimental research, 96 patients after PCI were randomly divided into the intervention group (49 cases), and the control group (47 cases) according to the discharge order. The intervention group received the individualized education model of cardiac rehabilitation for 12 weeks led by the cardiac rehabilitation outpatient nurse, and the control group received the routine follow-up guidance. Before and after the intervention of the two groups, the changes of cardiac rehabilitation knowledge were evaluated according to The Coronary Artery Disease Education Questionnaire-II(CADE-Q II), the compliance of cardiac rehabilitation exercise were evaluated according to heart rate band mobile APP or mobile WeChat exercise APP, the effects of cardiac rehabilitation exercise were adjusted according to a 6-minute walking test. Results: The increase scores of CADE-Q II in the intervention group after the intervention were higher than that in the control group:(10.04±2.75) points vs (5.57 ± 2.65) points, and the difference was significant (t=2.079, P < 0.05).The exercise compliance of daily aerobic exercise target rate indicated on the App of mobile phones in the intervention group were higher than that in the control group:88%(15/17) vs 53% (7/17), and the difference was significant (t=5.100, P <0. 05). The exercise compliance of daily exercises indicating on the Wechat application in the intervention group were higher than that in the control group: 82% (23/28) vs 48% (13/27), and the difference was significant (t=7.025, P <0.01).6MWD in the intervention group was higher than that in the control group: (48.77±26.67) mvs (16.77±43.65) m, and the difference was significant (t=3.534, P <0.01). Conclusion This cardiac rehabilitation education model can improve the knowledge level and compliance of cardiac rehabilitation exercise, and promote the cardiac rehabilitation of patients after PCI.

Objective: To explore the clinical effect and safety of minor liver resection for hilar cholangiocarcinoma (HC) of Bismuth-Corlette type Ⅲ and Ⅳ. Methods: From May 2007 to May 2017, the clinical data of 108 patients with Bismuth-Corlette type Ⅲ and Ⅳ HC underwent hepatectomy were collected and analyzed retrospectively.There were 56 males and 52 females, aged (57.2±5.3) years (ranged 48-76 years) .Among the 108 cases, there were 51 cases of type Ⅲa, 40 cases of type Ⅲb and 17 cases of type Ⅳ. Small-scale hepatectomy (≤3 hepatectomy) was performed in 70 cases, including 8 cases of 4b segment resection, 28 cases of 4b segment+5 segment resection, and 34 cases of partial 4 segment+partial 7 segment+partial 1 segment resection. Large-scale hepatectomy was performed in 38 cases (>3 segments) , of which 30 cases were treated with 2 segments+3 segments+4 segments+1 segment, and 8 cases were treated with 5 segments+7 segments+8 segments+1 segment. t′ test was used to analyze the data which did not conform to the normal distribution, and χ² test was used to calculate the incidence of postoperative complications and the 1, 3, and 5-year cumulative overall survival rate. Results: (1) The operation time of minor liver resection group ((180±25)minutes) was shorter than that of major liver resection group ((210±35)minutes) (t′=4.676, P<0.05) , the amount of blooding operation time of minor liver resection group ((310±80)ml) was less than that of major liver resection group ((500±110)ml)in the operation (t′=9.385, P<0.05) , and the difference was statistically significant. (2) The incidence of complications was lower in minor liver resection group and major liver resection group, and the difference was statistically significant (χ²=5.230, P<0.05) . (3) The actual 1-, 3- and 5-year survival rates were 87.1%, 58.4%, 30.0% and 84.2%, 57.9%,31.6%, respectively. There were no significant differences in survival rates in two groups in 1-, 3- and 5-year survival rates (χ²=0.177, P=0.674; χ²=0.005, P=0.946; χ²=0.029, P=0.865) . Conclusions Compared to patients with major liver resection, Minor liver resection for selected patients with HC of Bismuth-Corlette Ⅲ and Ⅳaccording to our criteria achieved better long-term outcomes. Chen′s biliojejunostomy is a simple, effective and safe method, which can be widely used when there are multiple biliary intestinal anastomosese.

Objective To investigate the relationship of blood concentration of tacrolimus with efficacy and safety in treatment of patients with myasthenia gravis.Methods The clinical data of 74 patients with myasthenia gravis admitted in our hospital from January 2013 to December 2015 were analyzed retrospectively.The blood concentrations of tacrolimus were determined by chemiluminescence microparticle immunoassay.Treatment effects were evaluated with clinical relative scoring in China.Results There were 41 men and 33 women in 74 patients with the mean age of (56.7 ± 15.8) years.According to the Osserman classification,there were 7 cases of type Ⅰ,18 cases of type Ⅱ b,27 cases of type Ⅱ a,5 cases of type Ⅲ and 17 cases of type Ⅳ.Forty seven patients underwent thymectomy,including 9 cases of thymoma and 38 cases of hyperplasia.Patients were followed up for (28.1 ± 14.5)days,and the effective rate was 74 ％ (55/74).There was no significant correlation between tacrolimus blood concentration and clinical efficacy.The incidence of adverse reaction of tacrolimus was 22％ (16/74),and the incidence of adverse reactions increased with the increase of blood concentration.Conclusion Tacrolimus is effective in treatment of myasthenia gravis.The correlation of blood concentration of tacrolimus with the clinical efficacy needs to be further studied,but certainly,the monitoring of blood concentration can reduce the incidence of adverse reactions.

Objective To investigate the effect of CYP3A5 genetic polymorphism on tacrolimus concentration in myasthenia gravis patients to explore the optimum doses for patients of different genotypes.Methods The CYP3A 5 genotypes of 95 myasthenia gravis patients treated with tacrolimus in our hospital from January 2013 to December 2015,were determined by fluorescence in situ hybridization and tacrolimus concentrations were determined by chemiluminescence microparticle immuno assay.The differences of tacrolimus concentrations/dosage (C/D) ratios,concentration and dosage oftacrolimus were compared among patients of different genotypes.Results There were 14 patients (14.74％) with CYP3A5*l*l,35 (36.84％) with CYP3A5*l*3 and46 (48.42％) with CYP3A5*3*3 in these 95 patients.The allele frequencies of *1 and *3 were 33.16％ and 66.84％,respectively.The C/D ratios of patients from the 3 groups were significantly different (F=24.860,P=0.000):the C/D ratio of CYP3A5*1/*1 patients was significantly lower than that of CYP3A5*1/*3 patients (P＜0.05),and that of CYP3A5*1/*3 carriers was significantly lower than that of CYP3A5*3/*3 patients (P＜0.05).The appropriate doses of tacrolimus of myasthenia gravis patients were different in different genotypes patients:the dose of patients with CYP3A5* 1* 1 was 4-5 mg/d,that of CYP3A5*3*3 was 2-3 mg/d,and that of CYP3A5*1*3 was between that of CYP3A5*1*1 and CYP3A5*3*3.Conclusions The polymorphism of CYP3A 5 genetic polymorphism has effect on plasma concentrations oftacrolimus in patients with myasthenia gravis.Clinical application ofpharmacogenetic studies will be helpful for individualization oftacrolimus appropriate dosage.

Objective To investigate the clinical value of amplitude integrated electroencephalogram on early diagnosis and prognosis evaluation of brain injury caused by neonatal asphyxia.Methods A total of 34 full-term asphyxiated neonates(asphyxia group)hospitalized in NICU of our hospital from January 2015 to September 2015 were selected;meanwhile,34 full-term healthy infants(control group)of the same term were selected.All cases were monitored for the activities of aEEG background,sleep-awakening cycle(SWC)and epileptic activity(SA)within 6 hours after birth.Meanwhile,the relationships between various indexes and asphyxia degree and brain injury were analyzed.Results The electroencephalogram of the asphyxia group was 52.9％and the rate of SWC was 58.8％,which were lower than those of the control group,and the difference had statistic significance(P<0.05).Meanwhile,neonates with epileptic activity in asphyxia group accounted for 11.8％,which was higher than that of control group significantly(P<0.05).Conclusion The AEEG changes of neonates at early period after birth are closely related to perinatal asphyxia and brain injury after asphyxia.The application of amplitude integrated electroencephalogram has an important significance on early diagnosis of neonatal asphyxia.

Objective: To evaluate the clinical efficacy of modified lateral window for maxillary sinus floor augmentation (MSFA). Methods: Fifty-five patients who visited the Stomatology Hospital Affiliated to Zhejiang University School of Medicine between June 2012 and October 2014 were enrolled in the study. Patients underwent MSFA with Bio-Oss grafts based on modified access window. During the operation the vertical height of the bony window was reduced from 6-8 mm of conventional oval window to 4-5 mm of slot-shaped window. The sinus membrane was detached completely via the lateral access and large particle Bio-Oss graft was placed in the sub-mucosal space. The implant survival, graft height, graft volume and resorption rates were measured. Intra-op and post-op complications were recorded. Results: There were 86 implants inserted. The 2-4 year cumulative survival rates were 97.67% by implant-based analysis and 96.36% by patient-based analysis. The residual bone height was (4.7±2.6) mm and bone width was (8.4±2.7) mm. The bone height of implantation site immediately after operation was (16.1±2.5) mm and it was (16.2±2.2) mm at restoration. The bone heights at 1 and 2 years after operation were (14.9±2.5) mm and (13.6±2.6) mm, respectively. The graft height was (10.6±2.8) mm and the graft volume was (1569±745) mm³ immediately after operation. The resorption rate of graft height 6 months after operation was 3.79% and that of graft volume was 7.87%. The 1-year accumulative resorption rate of graft height was 6.63% and that of graft volume was 10.89%. The 2-year accumulative resorption rate of graft height was 7.58% and that of graft volume was 15.26%. Small membrane perforation during MSFA was observed in 5 cases and all were successfully repaired by a collagen Bio-Gide membrane. Conclusion: The modified lateral technique obtains high implant survival rate, excellent graft stability and low complication rate at 2-4 year clinical follow-up, indicating that it is a safe, predictable and minimally invasive surgical method for severe atrophic maxillary posterior dentition.
Dental Implantation, Endosseous ; Follow-Up Studies ; Humans ; Maxillary Sinus ; surgery ; Sinus Floor Augmentation ; methods ; Treatment Outcome